Black liquor produced from pulping with a high value of chemical oxygen demand (COD) and biological oxygen demand (BOD) is highly harmful if discharged into the environment directly. One possible way to decrease the damage to the soil and water is to reuse the organic substances contained in it to cultivate yeasts for producing single-cell proteins (SCP) while reducing the COD. With this in mind, this study is devoted to treatment technology and the comprehensive utilization of black liquor. Various parameters were evaluated, and the COD of black liquor, initial pH, and nitrogen sources had significant influences on biomass and crude protein production. The research resulted in the maximum values of COD removal rate and crude protein production with 78.78 ±3.21% and 1.18 ±0.02 g/L achieved, respectively, under the optimized condition of black liquor concentration (60%), the recruitment of urea (0.5 g/L), initial pH (6.0), temperature (34 °C), shaking speed (180 rpm), and incubation time (36 h). Furthermore, this study provided a potential viable treatment of black liquor and revealed a feasible way to make full use of black liquor for the economical production of SCP.